Journal: Frontiers in Genetics

Article Title: Aberrant expression of the MID1 protein in neurons of Huntington’s disease brain

doi: 10.3389/fgene.2026.1753495

Figure Lengend Snippet: MID1 mRNA expression is aberrantly increased in HD-neurons. The MID1 expression was quantified in distinct cell populations isolated by MACS from the cortex of 15-week-old HdhQ150 knock-in mice (tg) and age-matched wild-type littermates (wt). The success of separation was quantified by qPCR analysis detecting GFAP as marker for astrocytes (A) , CD11b as marker for microglia (B) , and NeuN as neuronal marker (C) . (D) The marker proteins GFAP, NeuN, and CD11b were also detected on a Western blot of pooled samples from the tg animals (n = 7). MID1 was detected on the same blots. MID1 qPCR analysis revealed no statistically significant differences in (E) astrocytes ( p = 0.595) and (F) microglia ( p = 0.1966). (G) In neurons a significant upregulation of MID1 expression was detected (* p = 0.0104). MID1 levels were normalized to ACTB . Columns represent mean values ± SEM, p -values were calculated using an unpaired t-test with Welch’s correction (n wt = 7, n tg = 7).

Article Snippet: The Anti-ACSA-2 MicroBead Kit (130-097-678), the CD11b MicroBeads Kit (130-093-634), and the Neuron Isolation Kit (130-097-678) from Miltenyi Biotech were used for MACS.

Techniques: Expressing, Isolation, Knock-In, Marker, Western Blot

Journal: Bioactive Materials

Article Title: M2 macrophage-derived exosomes delivering haptoglobin and interleukin-10 plasmids for synergistic therapy of intracerebral hemorrhage

doi: 10.1016/j.bioactmat.2026.01.047

Figure Lengend Snippet: Schematic illustration of the preparation of M2-exo@HI and its mediated therapeutic mechanisms and signaling pathways. (a) RAW264.7 macrophages were polarized to M2 phenotype using DEX, followed by M2-exo isolation from cell supernatant via differential centrifugation. M2-exo@HI was prepared through encapsulating HI into M2-exo by electroporation. (b) M2-exo@HI crossed the BBB and localized to microglia in the hemorrhagic brain, delivering the HI plasmid into the nucleus. This prompted expression and secretion of Hp and IL-10 by microglia. The released Hp inhibited Hb toxicity by binding to Hb. IL-10 shifted microglial polarization from the pro-inflammatory M1 phenotype toward the reparative M2 phenotype, removing hematoma by engulfing erythrocyte and Hb-Hp complex, and decreasing pro-inflammatory cytokine levels—collectively enhancing neuroprotection and BBB repair. These beneficial outcomes were linked to the inhibition of the IL-17 and NF-κB signaling pathways and activation of the PI3K-Akt and ferroptosis pathways.

Article Snippet: For CLSM observation, 100 μL ICG and M2-exo@ICG were incubated with M1 microglia for 0.25 h, 0.5 h, 1 h and 2 h. Then cells were washed, fixed, stained with DAPI and observed by CLSM (A1R+, Nikon, Japan).

Techniques: Protein-Protein interactions, Isolation, Centrifugation, Electroporation, Plasmid Preparation, Expressing, Binding Assay, Inhibition, Activation Assay

Journal: Bioactive Materials

Article Title: M2 macrophage-derived exosomes delivering haptoglobin and interleukin-10 plasmids for synergistic therapy of intracerebral hemorrhage

doi: 10.1016/j.bioactmat.2026.01.047

Figure Lengend Snippet: Validation of M2-exo targeting, Hp/IL-10 transfection expression, and Hp/Hb binding. (a) Fluorescence imaging showing cellular uptake of ICG and M2-exo@ICG by M1 microglia. (b,c) Flow cytometry and corresponding quantification of RhB and M2-exo@RhB internalized by M1 microglia (n = 3). (d,e) Schematic illustration and quantitative analysis of the in vitro phagocytosis-release kinetics of M2-exo@RhB in BV2 under ICH-mimicking stimulation (n = 6). (f) Fluorescence images showing Hp and IL-10 expression in M1 microglia treated with M2-exo@HI for 12, 24, 48, 72 h. (g) Mean fluorescence intensity (MFI) quantification of Hp and IL-10 expression (n = 3). (h,i) ELISA measurements of secreted Hp and IL-10 protein levels (n = 3). (j,k) qPCR analysis of relative Hp and IL-10 mRNA expression (n = 3). (l) Western blot detection of Hp and IL-10 protein expression. (m) Densitometric quantification of Hp and IL-10 protein levels from Western blot (n = 3). (n) Co-immunoprecipitation assay confirming the formation of Hp-Hb complex. Data are presented as mean ± SD. Statistical significance was calculated by unpaired Student's t -test (c and e), and one-way ANOVA with Tukey's multiple comparisons test (g-k and m).

Article Snippet: For CLSM observation, 100 μL ICG and M2-exo@ICG were incubated with M1 microglia for 0.25 h, 0.5 h, 1 h and 2 h. Then cells were washed, fixed, stained with DAPI and observed by CLSM (A1R+, Nikon, Japan).

Techniques: Biomarker Discovery, Transfection, Expressing, Binding Assay, Fluorescence, Imaging, Flow Cytometry, In Vitro, Enzyme-linked Immunosorbent Assay, Western Blot, Co-Immunoprecipitation Assay

Journal: Bioactive Materials

Article Title: M2 macrophage-derived exosomes delivering haptoglobin and interleukin-10 plasmids for synergistic therapy of intracerebral hemorrhage

doi: 10.1016/j.bioactmat.2026.01.047

Figure Lengend Snippet: M2-exo@HI promotes in vitro microglia polarization, BBB repair and neuroprotection. (a) Flow cytometry analysis of M1-type (CD86 + ) and M2-type microglia (CD163 + ) following treatment with different formulations. (b,c) Percentages of CD86 + and CD163 + microglia populations (n = 3). (d–g) The cytokine levels of IL-10, TGF-β, TNF-α, and IL-1β in treated microglia (n = 3). (h) Fluorescence microscopy images showing erythrophagocytosis by microglia across treatment groups. (i) Schematic of the in vitro BBB model assessing FITC-dextran permeability using a transwell assay. (j) Quantitative analysis of FITC-dextran penetration (n = 7). (k) Flow cytometry analysis of neuronal apoptosis across treatments (n = 3). (l) Quantitative analysis of neuronal apoptosis (n = 3). Data are presented as mean ± SD. Statistical significance was tested by one-way ANOVA with Tukey's multiple comparisons test.

Article Snippet: For CLSM observation, 100 μL ICG and M2-exo@ICG were incubated with M1 microglia for 0.25 h, 0.5 h, 1 h and 2 h. Then cells were washed, fixed, stained with DAPI and observed by CLSM (A1R+, Nikon, Japan).

Techniques: In Vitro, Flow Cytometry, Fluorescence, Microscopy, Permeability, Transwell Assay

Journal: Bioactive Materials

Article Title: M2 macrophage-derived exosomes delivering haptoglobin and interleukin-10 plasmids for synergistic therapy of intracerebral hemorrhage

doi: 10.1016/j.bioactmat.2026.01.047

Figure Lengend Snippet: In vivo therapeutic effects of M2-exo@HI in hemorrhagic stroke. (a) The schematic diagram illustrates the construction of mouse cerebral hemorrhage model and treatment regimens. (b) Digital photos showing cerebral hematoma of ICH mice in different groups. (c) Quantitative measurements of hemoglobin concentration in different groups (n = 3). (d) Cerebral edema quantification by brain water content measurements (n = 3). (e) CLSM images showing M1 microglia (CD86 + , green) and M2 microglia (CD163 + , red) in different groups. Nucleus were stained with DAPI (blue). (f) Immunofluorescence staining showing co-localization of Hb/Hp with microglia in different groups. (g) Representative images of HE, Nissl, and TUNEL staining. Data are presented as mean ± SD. Statistical significance was tested by one-way ANOVA with Tukey's multiple comparisons test.

Article Snippet: For CLSM observation, 100 μL ICG and M2-exo@ICG were incubated with M1 microglia for 0.25 h, 0.5 h, 1 h and 2 h. Then cells were washed, fixed, stained with DAPI and observed by CLSM (A1R+, Nikon, Japan).

Techniques: In Vivo, Concentration Assay, Staining, Immunofluorescence, TUNEL Assay

Journal: bioRxiv

Article Title: Circulating extracellular vesicles drive microglial senescence and neurodegeneration in Parkinson’s disease

doi: 10.64898/2026.03.10.709299

Figure Lengend Snippet: (a) Immunostaining of HMC3 cells with DAPI (blue) and IBA1 (red) after 24 hours of EV treatment. Scale bar: 100 μm; zoom scale bar: 50 μm (b-c). Percentage and surface area of activated HMC3 cells after 24 hours of EV treatment. Data are represented as mean ± SEM and median with interquartile range (n=8 images per group). P-values are represented in the figures. (d) Heatmap showing normalized signal intensity of cytokine expression in HMC3 cells treated with PD-EV- and HC-EV . (e) Western blot quantification of p16 INK4a and p21 WAF /CIP in HMC3 cells after 48 and 96 hours of EV treatment (n=3). (f-g) Semi-quantification of p16 INK 4a and p21 WAF /CIP expressed as percentage change: (Normalized signal (EV-treated))/ (Normalized signal (PBS control)-1) *100. Data are represented as mean ± SEM. P-value is represented in the figures.

Article Snippet: Human microglia HMC3 cells (ATCC, Cat. No. CRL-3304) were maintained at 37 °C, 5% CO 2 in a DMEM/F-12 (Thermo Fisher Scientific) supplemented with GlutaMAX, 10% v/v FBS, 1% NEAA, and 1% P/S.

Techniques: Immunostaining, Expressing, Western Blot, Control

Journal: bioRxiv

Article Title: Circulating extracellular vesicles drive microglial senescence and neurodegeneration in Parkinson’s disease

doi: 10.64898/2026.03.10.709299

Figure Lengend Snippet: (a-c) p16 INK4a, p21 WAF /CIP , and SERPINE1 mRNA relative expression in HMC3 cells after 48 and 96 hours of treatment with HC-EV, PD-EV, or PBS.

Article Snippet: Human microglia HMC3 cells (ATCC, Cat. No. CRL-3304) were maintained at 37 °C, 5% CO 2 in a DMEM/F-12 (Thermo Fisher Scientific) supplemented with GlutaMAX, 10% v/v FBS, 1% NEAA, and 1% P/S.

Techniques: Expressing

Journal: bioRxiv

Article Title: Circulating extracellular vesicles drive microglial senescence and neurodegeneration in Parkinson’s disease

doi: 10.64898/2026.03.10.709299

Figure Lengend Snippet: (a) Schematic experimental setup of EV treatment on HMC3 cells followed by CM isolation and SH-SY5Y neurons treatment. (b,f) Immunostaining with DAPI (blue) and TUBB3 (red) of SH-SY5Y neurons after treatment with CM and EV - CM. Scale bar: 200 µm. (c,g) Neurite length quantification of SH-SY5Y neurons after 24 hours treatment with CM and EV - CM (n= 550-580 neurons per group). (d,h) Flow cytometry analysis of SH-SY5Y neurons exposed to CM and EV - CM for 24 hours. Annexin V FITC (x-axis) and propidium iodide (y-axis). Q1: dead cells (PI positive), Q2: late-stage apoptotic cells (Annexin V and PI positive), Q3: early-stage apoptotic cells (Annexin V positive), Q4: live cells (Annexin V and PI negative). (e,i) Bar graph showing percentage of cells in Q1 and Q2 stages after 24 hours treatment with CM and EV - CM. Results correspond to 3 replicates from at least three independent differentiations. (j) Neurite length and (k) apoptosis comparison of SH-SY5Y neurons treated with CM and EV - CM, respectively. Scatter plots show all data points, with median as a straight line and the IQR as an error bar. Bar plots show mean ± SEM. P-values are represented in the figures.

Article Snippet: Human microglia HMC3 cells (ATCC, Cat. No. CRL-3304) were maintained at 37 °C, 5% CO 2 in a DMEM/F-12 (Thermo Fisher Scientific) supplemented with GlutaMAX, 10% v/v FBS, 1% NEAA, and 1% P/S.

Techniques: Isolation, Immunostaining, Flow Cytometry, Comparison